Cultivation of Xylaria species biomass as a binding agent in material production

ABSTRACT

The process aseptically inoculates a liquid media with a vegetative Xylaria fungal species to form a culture; statically incubates the culture in a vessel for a time sufficient to begin initiation of fruit body development and asexual sporulation and halts incubation at maximum conidia production prior to the beginning of sexual sporulation. Thereafter, the entire culture contents of the incubation vessel are macerated to homogenize the fungal biomass and conidia therein and form an inoculum.

This application claims the benefit of U.S. Provisional PatentApplication No. 62/147,338, filed Apr. 14, 2015.

This invention relates to a process for generating fungal biomassthrough cultivation of species from the genus Xylaria. Moreparticularly, this invention relates to a process for generating fungalbiomass through cultivation of Xylaria polymorpha.

BACKGROUND

Xylaria species are generally saprophytic, soft rot ascomycetes thatprimarily grow on decaying hardwoods. During vegetative growth thisfungus extends white, monomitic mycelium to capture nutritionalresources. After external stimuli initiate fruiting, the outer myceliumsurface differentiates and forms black-pigmented stromatic tissue.Ascocarps (fruiting bodies) then elongate and extend upwards as theymature.

Fully mature fruiting bodies in the wild can be 3-10 cm tall and up to2.5 cm wide. These fungi have an extended fruiting cycle (3-4 months)consisting of an initial phase where diploid asexual conidia areproduced on the outer fruit body surface and a later phase whereperithecia eject haploid sexual ascospores. When in this younger stage,the asexual conidia-producing conidiophores are closely packed on thestroma surface.

Published US Patent Application 2014/0186,927 describes a process forthe production of a chlamydospore rich slurry inoculum for use ininoculating a solid or liquid substrate, such as described in publishedUS Patent Application 2008/0145,577. As is known in the prior art, aconcentrated mass of asexual spores can be used as an effectiveinoculum. Chiamydospores generated within normal vegetative growth canbe distributed and function as discrete points of inoculation. This isdependent upon the natural ability of the fungus to sporulateextensively in the mycelium. Such behavior is completely independent ofhyphal differentiation towards fruiting body development.

Spore mass inoculation has also been suggested in Published US PatentApplication 2005/0176583. The described method of spore production isreliant upon development of fully mature fruiting bodies on a solidsubstrate and submersing the fruiting bodies in water to capture thereleased sexual spores. The total process time from solid substrateinoculation to mature sporocarp harvest can take anywhere from 30-60days. The inoculum produced through this process is completely composedof sexual spores without incorporating mycelium or fruit body tissue.

OBJECTS OF THE INVENTION

Accordingly, it is an object of the invention to reduce the time forgenerating a biomass for use as an inoculum.

It is another object of the invention to reduce the time for generatinga biomass for use as a binding resin.

It is another object of the invention to streamline the process forgenerating a biomass for use as an inoculum or binding resin.

BRIEF SUMMARY OF THE INVENTION

Briefly, the invention provides a process for generating a biomasscomprising the steps of (1) preparing a liquid medium including a carbonsource, a nitrogen source and micronutrients; (2) inoculating the liquidmedia aseptically with a vegetative Xylaria fungal species to form aculture; (3) statically incubating the culture in a vessel for a timesufficient to begin initiation of fruit body development and asexualsporulation; (4) halting incubation at maximum conidia production priorto the beginning of sexual sporulation; and (5) thereafter maceratingthe entire culture contents of the incubation vessel to homogenize thefungal biomass and conidia therein and form an inoculum.

The behavior of Xylaria species allows for the development of a novelprocess that carries the benefits of asexual spore mass inoculation,while remaining reliant upon the growth of fruiting bodies and not uponsporulation within the mycelium.

It is understood that the process is particularly effective with avegetative Xylaria fungal species selected from the group consisting ofXylaria polymorpha, Xylaria hypoxylon, Xylaria filiformis and Xylarialongipes.

In accordance with the invention, a mass of tissue is generated onliquid media, which can then be induced to differentiate and producenumerous fruiting bodies within the same culture vessel. However, theascocarps do not need to reach full sexual maturity due to the primaryasexual sporulation phase. This reduces the total growth time frominoculation to ascocarp harvest from 60 days to 14-21 days.

The entire fungal mass can then be blended and diluted to function asinoculum.

The process of the invention incorporates the asexual conidia,vegetative hyphae, and fruit body tissue within the same inoculumproviding additional points of inoculation compared to the prior art. Asa means of generating large masses of fungal biomass with highbiological efficiency, this process greatly improves yield with a 41%increase in biomass compared to typical vegetative sheet biomassproduction.

The process of the invention can be used to make a fungal inoculum or afungal biomass for use as a binding resin.

In order to make a binding resin, the inoculum is combined withparticles, such as lignocellulosic particles, incubated for a timesufficient for hyphae to form a network around the particles and to bondthe particles into a cohesive biomass and thereafter the resultingcohesive biomass is set as a resin. In particular, the Xylaria speciesform dense networks around the individual particles (instead of bindingparticles together) as well as penetrate and form cavities within theparticle cells.

In making a fungal inoculum, the process is simplified by eliminatingthe need for isolation of spores from fruiting bodies or depending uponsporulation within the mycelium. All components of the tissue cultureare included in the inoculum (asexual conidia and mycelium), whichreduces processing time while potentially increasing the concentrationof discrete points of inoculation.

In making a binding resin, the process generates a fungal biomass persurface area of growth media that is 30% to 50% greater than typicalvegetative tissue cultures due to the prolific production of fruitingbodies with a homogenous stroma layer over the vegetative tissue layer.The binding resin can then be used with discrete particles or in thecasting of complex shapes.

These and other objects and advantages of the invention will become moreapparent from the following detailed description taken in conjunctionwith the accompanying drawings wherein:

FIG. 1 illustrates a flow diagram of a process for making an inoculum inaccordance with the invention;

FIG. 2 illustrates a flow diagram of a process for making a bindingresin biomass in accordance with the invention;

FIG. 3 illustrates a flow diagram of alternative steps in the process ofFIG. 2 in accordance with the invention;

FIGS. 4a and 4b pictorially illustrate views of developing ascocarpsafter 15 days of incubation;

FIG. 4c pictorially illustrates a view of isolated conidia after 15 daysof incubation;

FIGS. 5a, 5b and 5c pictorially illustrate views of elongated ascocarpsafter 45 days of incubation; and

FIG. 6 pictorially illustrates a view of a scanning electron micrograph(SEM) of a biomass made in accordance with invention after compression.

DETAILED DESCRIPTION

Referring to FIG. 1, the process for producing an inoculum requires thefollowing steps:

Step 1—Preparation of a Liquid Media.

A liquid growth medium is prepared by mixing:

-   -   a. Carbon Source: simple sugars, starch, malt extract    -   b. Nitrogen Source: Peptone, yeast extract, soy protein    -   c. Water

The amounts of the components are as follows:

-   -   Carbon Source: 8-15 g    -   Nitrogen Source: 0.5-4 g    -   Water: 1 L

After mixing of the media components, the mixture of components issterilized at 15 psi and 121° C. for 15 minutes.

Step 2—Inoculation of Liquid Media

The sterile liquid growth medium of step 1 is then inoculatedaseptically by combining with a previously prepared vegetative fungaltissue (I.e. mycelium) at a rate of 0.01-5% of mycelium to the sterileliquid growth medium (v/v=volume of inoculum added per unit volume ofmedia) via maceration to obtain a culture.

Step 3—Incubation of Culture

The inoculated culture of step 2 is incubated in a static culture vesselunder strict environmental conditions, i.e.

-   -   a. Growth temperature maintained between 20° C. and 30° C.    -   b. pH of media buffered to maintain 5-7 during growth    -   c. No exposure to light/UV    -   d. Oxygen/carbon dioxide exchange within the culture vessel        maintained to promote vegetative exponential growth        -   a. 12-22% Oxygen        -   b. 1-8% Carbon dioxide    -   e. Time: until vegetative growth develops a homogeneous mycelium        layer across the entire liquid surface, usually, between 5 and 7        days    -   f. Induction of ascocarp (fruiting body) development        -   a. Cycling of temperatures from 15° C. to 30° C. in 12 hour            periods to mimic day/night cycles. High temperatures            maintained during the “day” cycle and temperature is            gradually reduced for the “night” cycle.        -   b. Cycling of oxygen/carbon dioxide concentrations by            increasing gas exchange to maintain typical atmospheric            oxygen and carbon dioxide levels        -   c. Cycling of light/UV exposure from no light exposure to 6            to 18 hours of light exposure followed by a period of no            light exposure.

During this step, the inoculated culture is statically incubated (i.e.not stirred or agitated). Fruiting body development and asexualsporulation are initiated by altering the incubation environment once ahomogeneous mycelium layer develops across the entire liquid surface.

Incubation is halted at maximum conidia production prior to thebeginning of sexual sporulation. Maximum conidia production is indicatedwhen ascocarps drop a visible layer of conidia en masse from thefruiting bodies, usually between days 14 and 21.

Step 4—Harvesting of Biomass

After incubation of step 3, the entire culture contents of theincubation vessel are macerated to homogenize fungal biomass andconidia. Any remaining undigested sugars or extracellular growth factorsare maintained in the spent culture fluid and are carried through as apart of the inoculum.

Step 5—Verification of Conidia Production

The homogenized biomass from step 4 is analyzed for adequate asexualsporulation and vegetative hyphae development by taking a sample of thebiomass.

-   -   a. Verification of the presence of conidia is made via        microscopic features, i.e. conidia are observed via light        microscopy to verify diagnostic features, such as, size, shape,        texture, presence/absence of germ slit, color. A germ slit is an        opening in the spore cell wall where the germinating mycelium        emerges. This characteristic is only present in the sexual        ascospores and not the asexual conidia.    -   b. Concentration of conidia (spore concentration) is confirmed        via hemocytometer spore counts of culture liquid    -   c. Characterization of the hyphae types is made e.g. vegetative        hyphae are observed via light microscopy to determine physical        characteristics, such as, hyphae types, range of fragment        lengths, hyphal fragment density (concentration)    -   d. Determine total biomass present in the slurry.    -   e. Verify viability of both conidia and hyphae components using        viable colony forming units of combined biomass slurry and a        germination rate assay.

An assay of “viable colony forming units of combined biomass slurry” isa measure of discrete points of inoculation by incubating dilutions ofthe slurry liquid on a nutrient agar. The number of growing coloniesafter 24-72 hours indicates how many points of inoculation are presentper unit of slurry volume.

A germination rate assay determines what percentage of conidia areviable and germinate (begin germ tube extension from conidia) after24-48 hrs. This indicates the proportion of the inoculum that is viableas a result of conidia germination.

-   -   The assay can be conducted by different methods:

1. The first assay is a liquid culture incubated for 24 hrs and analyzedmicroscopically. The number of conidia forming germ tubes (germinating)are counted and the number of conidia not forming germ tubes(non-germinating) are counted until a total of at least 100 spores arecounted. This is repeated 3 times for each replicate bottle and anaverage percentage of germinating conidia is calculated(germinating/total spores×100%).

2. The second method is similar to the “viable colony forming units”assay. The slurry liquid is filtered through a 40 micrometer screen toeliminate fragments of mycelium and results in a suspension of conidia.Serial dilutions out to 0.00001% of the original solution are thenprepared and plated on nutrient agar. Plates are incubated for 24-72 hrsand the number of colonies is used to determine the number of viableconidia from the original sample conidia concentration.

Step 6—Qualification of Conidia/Hyphae Slurry for Use as an Inoculum

The conidia/hyphae slurry from the incubation vessel is combined withsterile water to dilute the biomass to an appropriate concentration forspecific applications.

-   -   a. Dilute to 0.1-5% for inoculation of a solid-state substrate.    -   b. Dilute to 0.01-5% for inoculation of liquid substrate, for        example, 1 part the biomass slurry and 99 parts water (a        100-fold dilution)

Referring to FIG. 2, the process for producing a biomass for use as abinding resin requires the following steps:

Step 1—Preparation of a Liquid Media.

A liquid growth medium is prepared as described above in step 1 of FIG.1.

Step 2—Inoculation of Liquid Media

The sterile liquid growth medium is inoculated as described above instep 2 of FIG. 1.

Step 3—Incubation of Culture

The inoculated culture is incubated in a suitable vessel under strictenvironmental conditions, i.e.

-   -   g. Growth temperature maintained between 20-30° C.    -   h. pH of media buffered to maintain 5-7 during growth    -   i. No exposure to light/UV    -   j. Oxygen/carbon dioxide exchange within the culture vessel        maintained to promote vegetative exponential growth        -   a. 12-22% Oxygen        -   b. 1-8% Carbon dioxide    -   k. Time: until vegetative growth develops a homogeneous mycelium        layer across the entire liquid surface, usually, between 5 and 7        days.

Step 4a—Induce Conidia Formation

The growth environment is altered to stimulate fruiting body (i.e.ascocarp) development and asexual sporulation phase, as above.

-   -   Cycling of temperatures    -   Cycling of oxygen/carbon dioxide concentrations    -   Cycling of light/UV exposure

Incubation is halted at maximum conidia production prior to thebeginning of sexual sporulation as in step 3 of FIG. 1.

Step 5a—Harvesting of Biomass

The entire culture contents of the incubation vessel are macerated tohomogenize fungal biomass and conidia as described above in step 4 ofFIG. 1.

Step 6a—Verification of Conidia Production

The homogenized biomass is analyzed as described above in step 5 of FIG.1.

Step 7a—Qualification of Conidia/Hyphae Slurry for Use as an Inoculum

The conidia/hyphae slurry from the incubation vessel is combined withsterile water as described above in step 6 of FIG. 1

Step 8a—Inoculation of Material Particles

The prepared conidia/hyphae slurry inoculum is aseptically combined withlignocellulosic particles in a mixing vessel to initiate fungalcolonization.

-   -   a. All lignocellulosic particle surfaces are covered with        conidia/hyphae slurry liquid to assure homogenous colonization        of substrate volume.

The diluted inoculum is added at 10-30% of the overall water addition,which translates to 2-10% of the total batch mass (particles+liquid).

Particle sizes can range from 0.125 inches to 0.75 inches.

Step 9a—Incubation of Inoculated Material Particles

The inoculated particles are incubated in a strictly controlledenvironment to optimize vegetative hyphae expansion, biomassaccumulation, and enzymatic activity.

-   -   a. Growth temperature maintained between 20-30° C.    -   b. 12-22% Oxygen    -   c. 1-8% Carbon dioxide    -   d. Limited exposure to light/UV

During this step, the temperature, pH, aeration and light exposure arecontrolled to maximize vegetative mycelium expansion for a timesufficient for hyphae to form a network around the particles and to forma cohesive biomass.

The environmental conditions are altered after the fungal biomass hashomogeneously covered all particle surfaces to elicit specificphysiological responses and to optimize fungal cell composition andexogenous enzyme activity.

The physiological responses can be inducement of pigmentation ormycelial surface morphologies for particular product aesthetics as wellas changing the cellular chemical makeup to provide strength increases.

The exogenous enzyme activity alters the characteristics of thelignocellulosic particles to improve physical characteristics of thefinal product such as strength, water swell, screw hold, and the like.

This step is dependent upon the targeted final product. Time ofcompletion of the step would be subject to appropriate quality controltesting per application.

Step 10a—Setting of Tissue Resin

The accumulated biomass is set as a resin using methods specific to thedesired product characteristics.

-   -   a. Heated convection drying, heated compression, cold        compression, freeze drying, microwave

Referring to FIG. 3, the process of FIG. 2 may be modified so that theparameters of the growth chamber may be maintained to maximize hyphalexponential growth and generate fungal biomass.

Thus, after step 3 (incubation of culture) of the process of FIG. 2, thefollowing steps are performed:

Step 4b—Induce Vegetative Hyphae Exponential Growth

-   -   a. Introduce nutrients optimized for biomass production as a        batch or fed-batch system.    -   b. Maintain optimum growth temperatures    -   c. Maintain dissolved oxygen levels with constant aeration of        liquid media    -   d. Strictly limit exposure of culture to light/UV    -   e. Induce specific target characteristics of fungal biomass via        staged nutrition addition and supplementation with induction        additives

During this step, the nutrition, temperature, aeration and lightexposure are optimized to maximize biomass yield and to induce thespecific target characteristics.

Step 5b—Harvest Biomass

The fungal biomass is separated from the spent culture media viafiltration.

-   -   a. Remove tissue biomass from spent media via filtration    -   b. Wash residual sugars, media components, and metabolites from        harvested biomass

Step 6b—Combine Fungal Biomass with Material Particles

The harvested biomass is mixed into the material particles tohomogenously cover particle surfaces as in FIG. 2, step 8a. However,there is no need to sterilize the contents of the mixing vessel.

Step 7b—Set Biological Resin

The biomass/particle mixture is set as a resin using methods specific tothe desired product characteristics.

-   -   a. Heated convection drying, heated compression, cold        compression, freeze drying, microwave

EXAMPLES

1. Production of Inoculum

-   -   a. Bottles with 500 mL of malt extract broth were sterilized and        aseptically inoculated by macerating half of a fully colonized        malt extract agar plate culture, i.e. a Xylaria polymorpha into        the media. The standard procedure is to use one full plate per        liter of liquid.    -   b. The bottles were shaken and placed at an angle to increase        media surface area. With the cap loosened, the bottles were        statically incubated at room temperature.    -   c. After 5 to 7 days, a tissue sheet formed on the surface of        the media and by day 15 the fruiting bodies had extended to        about 2 centimeters (cm) in height. In this case, it is an        important distinction that only half of the incubation time was        required for the formation of a homogeneous vegetative mycelium        sheet.        -   The tissue sheet from one bottle was then aseptically            transferred to a sterilized blender and macerated into 200            mL sterile water. This mixture was then poured back into the            bottle.    -   d. A sample of the mixture was aseptically removed and the spore        concentration was determined to be 0.38×10⁶ spores/mL using a        Hemocytometer.    -   e. This mixture was then used as the inoculum in Example 2.    -   f. The other bottles were allowed to continue incubating to 48        days after inoculation with the tissue sheet on the surface        producing fruiting bodies across more surface area.    -   g. The entire culture vessel contents were then aseptically        transferred to a sterilized blender and macerated to a        homogenous mixture.    -   h. A sample of the mixture was aseptically removed and the spore        concentration was determined to be 3.59×10⁶ spores/mL using a        Hemocytometer.

FIGS. 4a and 4b illustrate photomicrographs of the tissue sheet of stepc above after 15 days of incubation.

FIG. 4c illustrates a photomicrograph of isolated conidia from anascocarp sampled prior to the maceration in step c above.

FIGS. 5a, 5b and 5c illustrate photomicrographs of the tissue sheet ofstep f above after 45 days of incubation and prior to maceration. Theseimages show the continued development of the fruiting bodies and theincrease in conidia formation.

2. Agricultural Waste Inoculated with Inoculum Produced in Example 1

-   -   a. 500 g of corn stover as the agricultural waste and water were        combined in a 5 L filter patch bag and sterilized at 15 psi and        121° C. for 1 hour. 1300 mL of water was added to hydrate the        substrate to a suitable moisture content for facilitating        mycelial growth.    -   b. a 500 ml volume of the slurry prepared in Example 1 was then        added to an agricultural waste substrate after cooling to room        temperature to form a mixture.    -   c. The bag was agitated to distribute the inoculum and tools,        i.e. mold forms, were packed with 250 g of the substrate/slurry        mixture.    -   d. These tools were incubated at room temperature for 14 days        before parts were removed and convection dried.    -   e. Two one inch thick parts were stacked, heated and compressed        to 0.25 inches to set the tissue resin.    -   f. Mechanical strength as determined by the modulus of        elasticity and modulus of rupture of the material. was then        measured by using the standard 3-point bend test with an lnstron        Model 4411 and compared to an LD-1 Particleboard as defined by        ANSI A208.1-1999 Particleboard (1999).

Density Elastic Modulus Flexural Strength Part (lb/ft³) (psi) (psi)Xylaria sp. 38.66 157600 550 LD-1 Particleboard 35-40 79800 435

3. Biomass Production Comparison

-   -   a. Bottles of malt extract broth (500 mL each) were prepared and        sterilized at 15 psi and 121° C. for 1 hour.    -   b. One bottle was inoculated with a polyporaceae species with        aggressive vegetative growth and the other was inoculated with        Xylaria sp.    -   c. Inoculation was done aseptically by macerating half of a        colonized agar plate culture into the media broth.    -   d. Bottles were shaken and placed at an angle to increase broth        surface area. With the bottle cap loosened, incubation was        carried out statically at room temperature for 14 days.    -   e. At the end of incubation, tissue sheets were removed from        growth media and allowed to air dry to a constant mass.    -   f. Dry mass yields were compared and Xylaria sp. produced 41%        more dry mass than the polyporaceae species.

4 Tissue Added as Resin to Non-Colonized Particles

-   -   g. Inoculate liquid media by macerating vegetative fungal tissue        into the broth.    -   h. Statically incubate culture at room temperature in a vessel        with the maximum surface area per unit of broth volume.    -   i. Once numerous ascocarps form on the tissue sheet surface        (14-21 days), the tissue is removed from spent media and        homogenized in liquid.    -   j. The biomass slurry is then added to raw particles at a rate        of 12% dry tissue mass to dry particle mass.    -   k. Place mixture into mold of desired geometry.    -   l. Set tissue resin through drying or compression.

5. Methods to Set Tissue Resin

-   -   m. Static air drying (desiccation)    -   n. Heated drying (convection, microwave)    -   o. Freeze Drying    -   p. Heated Compression    -   q. Cold Compression

6. Casting Tissue Without Added Particles

-   -   r. Inoculate liquid media by macerating vegetative fungal tissue        into the broth.    -   s. Statically incubate culture at room temperature in a vessel        with the maximum surface area per unit of broth volume.    -   t. Once numerous ascocarps form on the tissue sheet surface        (14-21 days), the tissue is removed from spent media and        homogenized in liquid.    -   u. Place mixture into mold of desired geometry.    -   v. Set tissue resin through drying or compression.

7. White Finish Engineered Wood

-   -   w. Combine substrate particles to desired blend.    -   x. Add supplemental nutrition: clear flour, calcium sulfate,        spent brewers grain, algae waste, wheat bran, etc.    -   y. Add water to reach a moisture content of 60-70%.    -   z. Sterilize substrate blend at 15 psi and 121° C. for 1 hour.    -   aa. Dilute the inoculum prepared in Application A to 4.6×10⁵        spores/mL and mix into sterile substrate.    -   bb. Incubate at room temperature, low O₂, and high CO₂ for 7-14        days.    -   cc. Dry material to 5-10% moisture and compress with added heat        to a density of 40-60 lb/ft³.    -   dd. Can apply light-colored facing material without background        color affecting aesthetics.

8. Black Finish Engineered Wood

-   -   ee. Combine substrate particles to desired blend.    -   ff. Add supplemental nutrition: clear flour, calcium sulfate,        spent brewers grain, algae waste, wheat bran, etc.    -   gg. Add water to reach a moisture content of 60-70%.    -   hh. Sterilize substrate blend at 15 psi and 121° C. for 1 hour.    -   ii. Dilute the inoculum prepared in Application A to 4.6×10⁵        spores/mL and mix into sterile substrate.    -   jj. Incubate at room temperature with atmospheric O₂ and CO₂ for        7-14 days.    -   kk. Dry material to 5-10% moisture and compress with added heat        to a density of 40-60 lb/ft³.    -   ll. Black coloration can be aesthetically pleasing if no facing        material is needed.

What is claimed is:
 1. A process for generating a biomass comprising thesteps of preparing a liquid media including a carbon source, a nitrogensource and micronutrients; inoculating the liquid media aseptically witha vegetative Xylaria fungal species to form a culture; staticallyincubating the culture in a vessel for a time sufficient to inducefruiting body development and asexual sporulation; halting said step ofincubation at maximum conidia production prior to the beginning ofsexual sporulation; and thereafter macerating the entire culturecontents of the incubation vessel to homogenize the fungal biomass andconidia therein and form an inoculum.
 2. A process as set forth in claim1 wherein said step of statically incubating the culture includesmaintaining the culture at a temperature between 20° C. and 30° C. andat a pH of from 5 to
 7. 3. A process as set forth in claim 2 whereinsaid step of statically incubating the culture includes maintaining anoxygen/carbon dioxide exchange within the vessel to maintain an oxygenlevel from 12 to 22% oxygen and a carbon dioxide level of 1 to 8% carbondioxide.
 4. A process as set forth in claim 1 wherein said step ofinoculating the liquid media is at an amount of 0.01% to 5% m/v ofvegetative Xylaria fungal species to liquid media.
 5. A process as setforth in claim 1 wherein said vegetative Xylaria fungal species isselected from the group consisting of Xylaria polymorpha, Xylariahypoxylon, Xylaria filiformis and Xylaria longipes.
 6. A process as setforth in claim 1 wherein said step of statically incubating the cultureis conducted until vegetative growth develops a homogeneous myceliumlayer across an entire surface of said liquid media and continued withan altered incubation environment to induce fruit body development andasexual sporulation.
 7. A process as set forth in claim 6 wherein saidstep of statically incubating the culture is conducted for a time periodbetween 5 and 7 days until vegetative growth develops said homogeneousmycelium layer.
 8. A process as set forth in claim 6 wherein saidaltered incubation environment includes cycling of temperatures from 15°C. to 30° C. in 12 hour periods to mimic day/night cycles, cycling ofoxygen/carbon dioxide concentrations to maintain atmospheric oxygen andcarbon dioxide levels and cycling of light/UV exposure from no lightexposure to 6 to 18 hours of light exposure followed by a period of nolight exposure.
 9. A process as set forth in claim 1 further comprisingthe steps of combining the inoculum with lignocellulosic particles toinitiate fungal colonization of the particles; incubating the inoculatedmaterial particles for a time sufficient for hyphae to form a networkaround the particles and to form a cohesive biomass; thereafter settingthe biomass as a resin.
 10. A process as set forth in claim 9 whereinsaid particles are of a size of from 0.3 cm to 2 cm.
 11. A process asset forth in claim 9 wherein said step of incubating includesmaintaining the growth temperature between 20° C. and 30° C., an oxygenlevel of from 12% to 22%, a carbon dioxide level of from 1% to 8% and alimited exposure to light/UV.
 12. A process as set forth in claim 1further comprising the steps of combining the inoculum withlignocellulosic particles to initiate fungal colonization of theparticles; incubating the inoculated material particles for a timesufficient for hyphae to form a network around the particles and to forma cohesive biomass; removing the biomass from spent media; washingresidual media components from the biomass; combining the biomass withmaterial particles to initiate fungal colonization of the particles;incubating the combined biomass and material particles for a timesufficient for hyphae to form a network around the particles and to forma cohesive biomass; and thereafter setting the cohesive biomass as aresin.
 13. A process for generating a biomass comprising the steps ofpreparing a liquid media including a carbon source, a nitrogen source,micronutrients and a vegetative Xylaria fungal species to form aculture; statically incubating the culture in a vessel maintained in anincubation environment until vegetative growth develops a homogeneousmycelium layer across an entire surface of said liquid media in thevessel; thereafter altering said incubation environment to induce fruitbody development and asexual sporulation; thereafter halting said stepof incubation at maximum conidia production prior to the beginning ofsexual sporulation; and thereafter macerating the entire culturecontents of the incubation vessel to homogenize the fungal biomass andconidia therein and form an inoculum.
 14. A process as set forth inclaim 13 wherein said step of statically incubating the culture isconducted for a time period between 5 and 7 days until vegetative growthdevelops said homogeneous mycelium layer.
 15. A process as set forth inclaim 13 wherein said step of incubating includes maintaining the growthtemperature between 20° C. and 30° C., an oxygen level of from 12% to22%, a carbon dioxide level of from 1% to 8% and a limited exposure tolight/UV.
 16. A process as set forth in claim 15 wherein said step ofaltering said incubation environment includes cycling of temperaturesfrom 15° C. to 30° C. in 12 hour periods to mimic day/night cycles,cycling of oxygen/carbon dioxide concentrations to maintain atmosphericoxygen and carbon dioxide levels and cycling of light/UV exposure fromno light exposure to 6 to 18 hours of light exposure followed by aperiod of no light exposure.
 17. A process as set forth in claim 13wherein halting said step of incubation occurs in response to ascocarpsdropping a visible layer of conidia from the fruiting bodies.
 18. Aprocess as set forth in claim 17 wherein dropping a visible layer ofconidia from the fruiting bodies occurs between 14 and 21 days of saidstep of incubation.